title: Chitosan enhances transfection efficiency of cationic polypeptides/DNA complexes.
abstract: The aim of this research was to investigate the effect of cationic polypeptides mixed with chitosan (CS) on in vitro transfection efficiency and cytotoxicity in human cervical carcinoma cells (HeLa cells). The polypeptides/DNA complexes and ternary complexes (CS, polypeptides and DNA) at varying weight ratios were formulated and characterized by using gel electrophoresis. Their particle sizes and charge were evaluated. The effect of the type and molecular weight (MW) of polypeptides, the weight ratio, order of mixing, the pH and serum on transfection efficiency and cytotoxicity were evaluated in HeLa cells. Three types of polypeptides (poly-L-lysine; PLL, poly-L-arginine; PLA and poly-L-ornithine; PLO) were able to form complete complex with DNA at weight ratio above 0.1. The PLA MW >70 kDa showed the highest transfection efficiency. The order of mixing between CS, PLA and DNA affected the transfection efficiency. The highest transfection efficiency was observed in ternary complexes of PLA/DNA/CS (2:1:4) equal to PEI/DNA complex. For cytotoxicity studies, over 80% the average cell viabilities of the complexes were observed by MTT assay. This study suggests that the addition of CS to PLA/DNA is easy to prepare, safe and exhibits significantly improved DNA delivery potential in vitro.
authors.affiliation: Faculty of Pharmacy, Silpakorn University, Nakhon Pathom 73000, Thailand.
authors.forename: Praneet
authors.initials: P
authors.lastname: Opanasopit
authors.forename: Jintana
authors.initials: J
authors.lastname: Tragulpakseerojn
authors.forename: Auayporn
authors.initials: A
authors.lastname: Apirakaramwong
authors.forename: Tanasait
authors.initials: T
authors.lastname: Ngawhirunpat
authors.forename: Theerasak
authors.initials: T
authors.lastname: Rojanarata
mesh_terms: D002412:Cations; D002470:Cell Survival / Q000235:genetics; D048271:Chitosan / Q000737:chemistry / Q000494:pharmacology*; D004247:DNA / Q000008:administration & dosage*; D004587:Electrophoresis, Agar Gel; D005260:Female; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D010316:Particle Size; D010455:Peptides / Q000737:chemistry*; D011107:Polylysine / Q000737:chemistry; D014162:Transfection / Q000379:methods*; D002583:Uterine Cervical Neoplasms / Q000235:genetics / Q000473:pathology
chemical_list: D002412:Cations; D010455:Peptides; C008973:polyornithine; D011107:Polylysine; C015462:polyarginine; D004247:DNA; D048271:Chitosan